Protease were produced by Trichoderma harzianum and Talaromyces flavus when its were grown in the presence of casein. Proteases from the culture filtrate of T. harzianum and T. flavus were purified and tested for their antifungal activity against brown spot disease caused by Botrytis fabae on faba bean. The enzymes were purified in a 2-step procedure involving ammonium sulfate precipitation and Sephadex G-200 gel permeation chromatography. Both enzymes have an optimum temperature of 30 and optimal pH value of 6.5. T. flavus exhibited high levels of extracellular protease activity compared with T. harzianum. Germination and growth rate, extracellular polygalacturonase (PGase) and carboxymethyl cellulase (CMCase) activities of Botrytis fabae were inhibited by the purified protease at a concentration of 40-120 U/ml. Growth and extracellular production of B. fabae were completely inhibited by the protease enzyme of T. flavus at a concentration of 80 U/ml, while protease from T. harzianum was effective at 120 U/ml. Proteases were effective in reducing brown spot disease severity and pathogen sporulation on faba bean leaves inoculated with B. fabae. Proteases at a low concentration of 80 U/ml inhibited B. fabae on leaf surface, as determined by Scanning Electron Microscope (SEM) examination. When enzymes were applied as a foliar spray on faba bean plants grown under natural infested field conditions, the disease incidence was greatly reduced. Protease from T. flavus at 80 U/ml was more significant in reducing disease incidence on leaves and pod and increased pod yield/plant. These results indicated that protease can be a safe biodegradable biocontrol agent for control of brown spot disease on fabae bean.